CN107202887A - The detection method of the related specific antigen of the information-based tumour of rapid qualitative a kind of, detection means and preparation method thereof - Google Patents
The detection method of the related specific antigen of the information-based tumour of rapid qualitative a kind of, detection means and preparation method thereof Download PDFInfo
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Abstract
The detection means of the invention that a kind of related specific antigen of the information-based tumour of rapid qualitative is provided, including the related specific antigen detector bar of tumour and instrument, network transmission device and terminal count device.Wherein, the reaction zone is provided with detection zone, and the detection zone is provided with detection line T and nature controlling line C.Add and " solid-phase coating labelled antibody-determined antigen-labelled antibody " immune complex is formed after sample, its color intensity is directly proportional to labelled antigen concentration to be measured.Such as 10-15 minutes particular detection time, chromatography detector bar (box) is detected by an unaided eye to record result or transmitting to the terminal count device by network transmission device are read in result or quantitative analysis instrument.Such as by relevant information by computer or any network equipment, or smart mobile phone, storage sends person under test, such as operating personnel, doctor, or be sent to inspection center to.
Description
Technical field
The invention belongs to biomedical inspection field, more particularly to a kind of information-based tumour of rapid qualitative are related special anti-
Former detection method, detection means and preparation method thereof.
Background technology
According to epidemiological statistics, tumor patient has hundreds thousand of or even up to a million patients every year, and numeral is all on constantly
Rise.Current screening method is enzyme-linked method or luminescence method, PCR, CT, or MRI.Program is complicated, needs professional equipment and professional to grasp
Make.Also there is a small amount of flash chromatography method in the market, for example, detect prostate cancer and non-specific Tumor invasion CEA, but only
Energy qualitative detection, it is impossible to make a definite diagnosis and monitor follow-up.Other main tumour specific antigen quick detections, sensitivity extreme difference is also
Qualitative range estimation, no value for clinical application.
The content of the invention
In view of this, it is an object of the invention to provide a kind of inspection of the related specific antigen of the information-based tumour of rapid qualitative
Survey device, including the related specific antigen detector bar of tumour and instrument, network transmission device and terminal count device.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
Tumour correlation specific antigen detector bar includes sample uptake zone, filtering area, chromatographic zone and suction zones.
Preferably, the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention includes bottom
Plate, sample uptake zone (bar), filtering area (bar), chromatographic zone (bar) and suction zones (bar) are pasted onto on viscose glue bottom plate, are assembled
Bottom plate；The present invention is preferably first from bottom to top to paste sample uptake zone (the 1st area), filtering area successively in bottom plate to the assembling
(the 2nd area), chromatographic zone (the 3rd area), suction zones (the 7th area), paste make on viscose board, during stickup each area overlapping 1~
1.5mm；In the present invention, the width of the slitting is preferably 3.0~5.0mm, preferably 4mm.
The present invention does not have special limitation to the material, size and source of the bottom plate, ripe using those skilled in the art
The bottom plate being used in immunoassay strip known.In the present invention, the material of the bottom plate is preferably plastics or film condensation material.
After the bottom plate assembled, the bottom plate of assembling is cut, immunity test strip is obtained.The present invention does not have to the method for the slitting
Have specifically limited, using slitting method well-known to those skilled in the art.
The immunoassay strip that the present invention is provided includes the sample uptake zone being arranged on the bottom plate.In the present invention, institute
State the material preferably glass fibre of sample uptake zone.The present invention does not have any limitation, ability to the source of the glass fibre
Glass fibre known to field technique personnel.The present invention is pasted to the sample uptake zone in the lower end of bottom plate, will be described
The lower edge of sample uptake zone is concordant with bottom plate lower edge.
In the present invention, the sample that the sample uptake zone absorbs is blood, serum or blood plasma.The source of the blood does not have
Limitation, using blood well-known to those skilled in the art or serum, blood plasma.The sample added in the embodiment of the present invention
Can be finger blood.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
Chromatographic zone includes immobilized label antibody district, detection zone and check plot；The solid phase labelling antibody district is arranged on the bottom plate,
Above sample uptake zone, and solid phase labelling antibody district 1.5mm overlapping with sample uptake zone.In the present invention,
The material of the solid phase labelling antibody district is preferably glass fibre.
Preferably, in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention, also wrap
The reaction zone being arranged on the bottom plate is included, the reaction zone is close to the solid phase labelling antibody district, positioned at the solid phase labelling
The top of antibody district, and the reaction zone and the solid phase labelling antibody area overlapping 1.5mm.In the present invention, the reaction
The material in area is preferably nitrocellulose filter.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
Reaction zone is provided with detection zone, and the detection zone is provided with detection line T and nature controlling line C.The detection line T is located at nature controlling line C's
Below, the detection line T is close to solid phase labelling antibody district, and the nature controlling line C is close to suction zones.
The detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention is to be based on following principle：
Sample is blood sample：Blood, or refer to blood, or serum, or blood plasma.Sample application zone (the 1st area) is added drop-wise to, blood sample need not be located
Reason, borrows or does not go upward to red blood cell blood cell in filtering area (the 2nd area), blood sample by developping solution and be stranded in filtering area, the liquid of filtration
The labelled antibody of body dissolving solid phase labelling antibody district (the 4th area) enters chromatographic zone (the 3rd area).Such as there is antigen in blood sample, then with
Labelled antibody formation " antigen-labelled antibody " compound.The compound is up on chromatography strip by capillary theory, to the 5th area
On detection zone, another specific mark antibody formation " labelled antibody-antigen-solid phase labelling antibody " with one, T areas solidus
Composite precipitation line.The color of the precipitation line will be in direct ratio with concentration in intensity antigen blood sample.Double antibody sandwich method principle-in T
Detection zone formation " solid-phase coating labelled antibody-determined antigen-labelled antibody " immune complex, its color intensity is with treating mark
Note antigen concentration is directly proportional.Such as 10-15 minutes particular detection time, chromatography detector bar (box) is detected by an unaided eye and records knot
Result is read in fruit or quantitative analysis instrument.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
Instrument is handheld portable detector.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
Handheld portable detector is made up of the readout instrument based on smart mobile phone, inspection software, cloud service.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
The transmission means of network transmission device includes the one or more of following methods：
By the cloud service, through wechat downloading data；
Via the detector output software, through USB output datas；
Printed out from wechat or bluetooth printing machine；
Sent from the readout instrument by lettergram mode.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
The related specific antigen concentration of tumour of testing sample and the face of detector have been stored in the inspection software of handheld portable detector
Linear relationship curve between intensity of colour.
Preferably, it is described in the detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention
It is special that tumour correlation specific antigen includes mammary tumor specific antigen CA-125, mammary tumor specific antigen CA-153, mammary tumor
Hapten CA-199, ED-SCLC specific antigen proGRP, non-small cell lung cancer specific antigen Cyfra21-1 one kind or several
Kind.
Another aspect of the present invention is to provide a kind of detection method of the related specific antigen of the information-based tumour of rapid qualitative,
Comprise the following steps：
1) sample (blood, or serum, or blood plasma) is added dropwise to the sample pipetting volume area of above-mentioned specific antigen detector bar；
2) sample by or not by developping solution, into above-mentioned filtering area, chromatographic zone；
If 3) there is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line in chromatographic zone, illustrate detection knot
Fruit is the positive, if there is not the composite precipitation line, illustrates to be detected as feminine gender；When the sample enters check plot,
Control line should occur, when such as there is no control line, detection failure；
4) " labelled antibody-antigen-solid-phase coating antibody " the composite precipitation line is carried out using above-mentioned instrument
Color intensity is determined, and is obtained antigen concentration in sample, is transmitted by above-mentioned network transmission device to the terminal count device.
Another object of the present invention is to provide the detection dress of the related specific antigen of the information-based tumour of above-mentioned rapid qualitative
The preparation method put, comprises the following steps：
1) labelled antibody is dissolved in buffer solution, the labelled antibody diluted, by the labelled antibody of described dilution
Glass fibre membrane is sprayed on, is then dried in vacuum desiccator, solid phase labelling antibody membrane is obtained；
2) coated antibody solution is lined and detection line is formed on reaction zone, rabbit anti-mouse igg solution is lined into reaction zone
Upper making nature controlling line formation check plot, the nitrocellulose film after line is placed under 37 DEG C of environment and dried 25 minutes, is coated with
Labelled antibody chromatographic film；
3) will be first in bottom plate from bottom to top paste sample uptake zone (the 1st area), filtering area (the 2nd area), chromatographic zone (the successively
3rd area), suction zones (the 7th area), paste makes the bottom that the overlapping 1~1.5mm in each area is assembled on viscose board, during stickup
After plate, the bottom plate of assembling is cut, the related specific antigen detector bar of tumour is obtained；
4) configuration instrument, network transmission device and terminal count device.
Compared with prior art, the present invention has advantages below to the present invention：
1. it is specific high：Required pairing labelled antibody acts only on the related specific antigen of high specific detection tumour.
2. sensitivity is high：Required pairing labelled antibody is with the related specific antigen of high-sensitivity detection tumour, with enzyme linked immunosorbent assay, hair
Light method is close, can reach 0.1ng/ml detection level.
3. related specific antigen trial zone (T) shade of detection box detection tumour or survey that required pairing antibody is fabricated to
Line strength specific antigen concentration related to tumour is tried linear, by the detector and network transmission that are stored in cloud service
Device, can easily obtain quantitative testing result.
4. can a step quick detection, blood sample to be measured need to be only added dropwise in the detection box that required pairing labelled antibody is fabricated to,
Just result can be measured within 10-15 minutes.
5. testing result quantitative information, by the way that the related specific antigen detector bar of tumour is inserted into detecting instrument, enters at once
Enter digitization procedure to show, and can be by relevant information by computer or any network equipment, or smart mobile phone, storage, which is sent to, to be treated
Survey person, such as operating personnel, doctor, or it is sent to inspection center.
Brief description of the drawings
Fig. 1 is the related specific antigen detector bar structural representation of the information-based tumour of rapid qualitative of the present invention；
Fig. 2 is the testing result schematic diagram of individual event tumour specific antigen detector bar in the embodiment of the present invention；
Fig. 3 is the testing result schematic diagram of individual event tumour specific antigen detection box in the embodiment of the present invention；
Fig. 4 is the testing result schematic diagram of double item tumour specific antigens detection boxes in the embodiment of the present invention；；
Fig. 5 is the tumour specific antigen detection means schematic appearance in the embodiment of the present invention；
Fig. 6 is signal schematic diagram in smart mobile phone position in the tumour specific antigen detection means in the embodiment of the present invention；
Fig. 7 shows for quantitative information result output display in the tumour specific antigen detection means in the embodiment of the present invention
It is intended to.
Embodiment
An embodiment provides a kind of detection dress of the related specific antigen of the information-based tumour of rapid qualitative
Put, including the related specific antigen detector bar of tumour and instrument, network transmission device and terminal count device.
In one embodiment of the invention, the related specific antigen detector bar of the tumour has following structure, such as Fig. 1 institutes
Show,：
1st area is sample uptake zone (sample application zone),
2nd area is filtering area,
3rd area is chromatographic zone (nitrocellulose chromatographic film),
4th area immobilized label antibody district A,
5th area is detection zone T, including labelled antibody spraying area band,
6th area is check plot C,
7th area is suction zones,
Wherein, the chromatographic zone includes immobilized label antibody district, detection zone and check plot.
In one embodiment of the invention, the information-based tumour of rapid qualitative of the present invention related specific antigen
The sample uptake zone (bar) of detection means, filtering area (bar), chromatographic zone (bar) and suction zones (bar) are arranged on bottom plate；The present invention
It is preferably first from bottom to top to paste sample uptake zone (the 1st area), filtering area (the 2nd area), chromatography successively in bottom plate to the assembling
Area (the 3rd area), suction zones (the 7th area), paste make the overlapping 1~1.5mm in each area on viscose board, during stickup；The present invention
In, the width of the slitting is preferably 3.0~5.0mm, preferably 4mm.
The present invention does not have special limitation to the material, size and source of the bottom plate, ripe using those skilled in the art
The bottom plate being used in immunoassay strip known.In the present invention, the material of the bottom plate is preferably plastics or film condensation material.
After the bottom plate assembled, the bottom plate of assembling is cut, immunity test strip is obtained.The present invention does not have to the method for the slitting
Have specifically limited, using slitting method well-known to those skilled in the art.
The immunoassay strip that the present invention is provided includes the sample uptake zone being arranged on the bottom plate.In the present invention, institute
State the material preferably glass fibre of sample uptake zone.The present invention does not have any limitation, ability to the source of the glass fibre
Glass fibre known to field technique personnel.The present invention is pasted to the sample uptake zone in the lower end of bottom plate, will be described
The lower edge of sample uptake zone is concordant with bottom plate lower edge.
In the present invention, the sample that the sample uptake zone absorbs is blood, serum or blood plasma.The source of the blood does not have
Limitation, using blood well-known to those skilled in the art or serum, blood plasma.The sample added in the embodiment of the present invention
Can be finger blood.
In one embodiment of the invention, the solid phase labelling antibody district is arranged on the bottom plate, is inhaled positioned at sample
Receive above area, and solid phase labelling antibody district 1.5mm overlapping with sample uptake zone.In one embodiment of the present of invention
In, the material of the solid phase labelling antibody district is preferably glass fibre.
In one embodiment of the invention, the reaction zone is close to the solid phase labelling antibody district, positioned at the solid phase
The top in labelled antibody area, and the reaction zone and the solid phase labelling antibody area overlapping 1.5mm.In the reality of the present invention
Apply in example, the material of the reaction zone is preferably nitrocellulose filter.
In one embodiment of the invention, the reaction zone is provided with detection zone, and the detection zone is provided with detection line T
With nature controlling line C.The detection line T is located at below nature controlling line C, and the detection line T is close to solid phase labelling antibody district, the Quality Control
Line C is close to suction zones.
In one embodiment of the invention there is provided the preparation method of the detector bar of the related specific antigen of tumour, including
The following steps：
Labelled antibody is added into colloid gold label solution, after stirring 10~15 minutes, the calf serum that concentration is 1% is added
Protein liquid, the Bovine serum albumin liquid is 1 with colloid gold label liquor capacity ratio：7~15, centrifuged through (12000PM) at a high speed
After ten minutes, colored precipitate thing is obtained, the colored precipitate thing is labelled antibody.Serum solution in extraction, sediment is dissolved in spy
Determine in buffer solution.
The present invention is not particularly limited to the source of the colloid gold label solution, using well known to those skilled in the art
Colloid gold label solution.
In the present invention, the centrifugal rotational speed is preferably 12000vpm, and the temperature of the centrifugation is 4 DEG C.
In the present invention, the method for the labelled antibody is not particularly limited, dilution side well known to those skilled in the art
Method.
Obtain after labelled antibody, mark is sprayed on glass fibre membrane.The spraying is preferably immersion, hand coatings or instrument
Device is sprayed, using the method for immersion in the embodiment of the present invention.Labelled antibody is then by the tunica fibrosa after immersion in vacuum desiccator
Under, it is dried in vacuo, obtains solid phase labelling antibody district.In the present invention, the temperature of the drying is preferably 35~38 DEG C, 8
Solid phase labelling antibody membrane is made in or so hour.
The preparation of coated antibody：
The vacuum drying of labelled antibody solution is lined into making detection line T lines on nitrocellulose film (the 5th area) reaction zone, will
Rabbit anti-mouse igg solution, which is lined, makes nature controlling line C line (the 6th area), the check plot rule on reaction zone.The present invention is to described
The mode of line is not restricted, using labelled antibody spraying method well known to those skilled in the art.The present invention is implemented
Line is rule using lining instrument in example, model Biojet, the Biodot CA, USA of the lining instrument.In the present invention one
In individual embodiment, the concentration of the coating buffer is preferably 2mg/ml；The coating buffer quantity for spray is preferably per 30cm nitrocelluloses
Film is sprayed 80ul/ seconds
After line, nitrocellulose film is placed under 37 DEG C of environment and dried 25 minutes by the present invention, obtains coating labelled antibody layer
Analyse film.The present invention is not particularly limited to the method for the drying, is using furnace drying method well-known to those skilled in the art
Can.In an embodiment of the invention, sample absorbing strip, filtering rod, mark chromatography strip and water suction bar are pasted onto viscose glue bottom plate
On, the bottom plate assembled.In an embodiment of the invention, it is preferably first from bottom to top to be glued successively in bottom plate to the assembling
Note sample uptake zone (the 1st area), filtering area (the 2nd area), chromatographic zone (the 3rd area), suction zones (the 7th area), are pasted on viscose board,
Make the overlapping 1~1.5mm in each area during stickup.
After the bottom plate assembled, the bottom plate of assembling is cut, the detector bar of the related specific antigen of tumour is obtained.This
Invention is not particularly limited to the method for the slitting, using slitting method well-known to those skilled in the art.This hair
Slitting is using cutting machine slitting, the cutting machine model K in bright embodiment:nematic2360CA,USA.Of the invention one
In embodiment, the width of the slitting is preferably 3.0~5.0mm, preferably 4mm.
In an embodiment of the invention, described antibody labeling particle can be collaurum, and magnetic particle, carbon particle shines grain,
Glue gold body grain, silver granuel, colored latex grain, fluorescence, textile dyestuff, enzyme, quantum dot, liposome, other particles etc..In the present invention one
In individual embodiment, the described preferred collaurum of antibody labeling particle.
In an embodiment of the invention, the related specific antigen of described tumour is preferably ovarian neoplasm specific antigen
CA125, ovarian neoplasm specific antigen CA153, ovarian neoplasm specific antigen CA199, ED-SCLC antigen gastrin releasing peptide
Precursor ProGastrin Releasing Peptide, referred to as (ProGRP)；Non-small cell lung cancer antigen cyokeratin antigen
Cyffa 21-1。
In embodiments of the invention, the antibody sources of the corresponding antibody of the tumor markers and labelled antibody are all from
JAJ international, Inc.San Diego, CA, USA.Tumour specific antigen described in the embodiment of the present invention
The product type of coated antibody and labelled antibody see the table below 1.
The tumour of table 1 correlation specific antigen pairing labelled antibody product type title
| Tumour correlation specific antigen | Detection zone (T) coated antibody | Labelled antibody |
| CA125 | #125-4012 | #125-3401 |
| CA153 | #153-782 | #153-678 |
| CA199 | #199-1033 | #199-1214 |
| Pro-GRP | #GRP-084 | #GRP-086 |
| Cyfra | #Cyf-124 | #Cyf-138 |
The detection means of the related specific antigen of the information-based tumour of rapid qualitative of the present invention is to be based on following principle：
Sample is blood sample：Blood, or refer to blood, or serum, or blood plasma.Sample application zone (the 1st area) is added drop-wise to, blood sample need not be located
Reason, borrows or does not go upward to red blood cell blood cell in filtering area (the 2nd area), blood sample by developping solution and be stranded in filtering area, the liquid of filtration
The labelled antibody of body dissolving solid phase labelling antibody district (the 4th area) enters chromatographic zone (the 3rd area).Such as there is antigen in blood sample, then with
Labelled antibody formation " antigen-labelled antibody " compound.The compound is up on chromatography strip by capillary theory, to the 5th area
On detection zone, another specific mark antibody formation " labelled antibody-antigen-solid phase labelling antibody " with one, T areas solidus is multiple
Close precipitation line.The color of the precipitation line will be in direct ratio with concentration in intensity antigen blood sample.Double antibody sandwich method principle-in T inspections
Survey area and form " solid-phase coating labelled antibody-determined antigen-labelled antibody " immune complex, its color intensity and mark to be measured are anti-
Original content is directly proportional.In particular detection time such as 10-15 minutes, chromatography detector bar (box) is detected by an unaided eye and records result or fixed
Result is read in amount analyzer.
Preferably, in one embodiment of the invention, the instrument is handheld portable detector.
Preferably, in one embodiment of the invention, the handheld portable detector is based on smart mobile phone
Readout instrument, inspection software, cloud service composition.
Preferably, in one embodiment of the invention, the transmission means of the network transmission device includes following methods
One or more：
By the cloud service, through wechat downloading data；
Via the detector output software, through USB output datas；
Printed out from wechat or bluetooth printing machine；
Sent from the readout instrument by lettergram mode.
In an embodiment of the present invention, quantitative information readout instrument used is QikTech-4000 (JAJ
International, Inc.San Diego, CA, USA), it is a kind of image immunoassays complete equipment, by " in smart mobile phone
Based on readout instrument；Inspection software；Cloud service " is constituted.
QikTech-4000 is handheld portable detector, its outward appearance as shown in figure 5, will detect box (detector bar when using
Outer layer sets the detection box of plastic housing) insertion detector, based on intelligent quick diagnosis test (RDT) reader, carry out image and adopt
Collection, processing and analysis.
Detect the color intensity relation proportional to test substance of detection line in box.This color or luminous intensity are color
Intensity is marked such as gold, or is fluorescence intensity, or is luminous intensity.Color intensity readout instrument can be read to the first image of testing result
As a result, then Digitization Software is analyzed.Most a whole set of result is inputted to high in the clouds at last.Once will detection box insertion readout instrument, intelligence
The open lighting device of mobile phone hardware communication, this lighting device (smart mobile phone and its connection with other parts below detection box
Relation is as shown in Figure 6), this narrow LED light, through detection box, detects detection line (T) color intensity, passes through intelligence
Mobile phone enters digital shadow record program.Intensity to be measured is made comparisons with the standard curve (see embodiment) stored in software, is directly shown
The concentration of test substance is shown：Units per ml, the testing result of quantitative information is as shown in Figure 7.
Preferably, in one embodiment of the invention, stored in the inspection software of the handheld portable detector
Linear relationship curve between the related specific antigen concentration of the tumour of testing sample and the color intensity of detector.
Preferably, in one embodiment of the invention, the related specific antigen of the tumour specifically resists including mammary tumor
Former CA-125, mammary tumor specific antigen CA-153, mammary tumor specific antigen CA-199, ED-SCLC specific antigen
ProGRP, non-small cell lung cancer specific antigen Cyfra21-1 one or more.
There is provided a kind of inspection of the related specific antigen of the information-based tumour of rapid qualitative in one embodiment of the invention
Survey method, comprises the following steps：
1) sample that sample (blood, or serum, or blood plasma) is added dropwise into above-mentioned specific antigen detector bar adds
Sample area；
2) sample by or not by developping solution, into above-mentioned filtering area, chromatographic zone；
If 3) as shown in Fig. 2 occurring " labelled antibody-antigen-solid-phase coating antibody " in chromatographic zone again
Precipitation line is closed, then it is the positive to illustrate testing result, if there is not the composite precipitation line,
Illustrate to be detected as feminine gender；When the sample enters check plot, control line (shown in Fig. 2) should occur,
When such as there is no control line, detection failure；
4) " labelled antibody-antigen-solid-phase coating antibody " the composite precipitation line is carried out using above-mentioned instrument
Color intensity is determined, and is obtained antigen concentration in sample, is transmitted by above-mentioned network transmission device to the terminal count device.
Preferably, in another embodiment of the present invention, the tumour specific antigen detector bar outer layer can be provided with
Plastic casing, as shown in figure 3, the result for a kind of individual event (only having tumour specific antigen in sample) detection box (detector bar) is shown
It is intended to, wherein, there is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line T, then it is the positive to illustrate testing result, such as
There is not the composite precipitation line (T) in fruit, then explanation is detected as feminine gender；When the sample enters check plot, should occur pair
According to line C (shown in Fig. 3), when such as there is no control line, then detection failure.
Preferably, in another embodiment of the present invention, the tumour specific antigen detector bar can test two kinds or
Two or more tumour specific antigen, as shown in figure 4, being double items (only having two kinds of tumour specific antigens in sample) detection box
The result schematic diagram of (detector bar), wherein, there is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line T1, T2, then
Illustrate that testing result is tied for two kinds of antigens of tumour-specific (such as ovarian neoplasm specific antigen CA125, CA153) to be positive
Really, if there is not the composite precipitation line (T), illustrate to be detected as feminine gender；When the sample enters check plot, it should go out
Existing control line C (shown in Fig. 3), when such as there is no control line, then detection failure.
Preferably, it is related special anti-there is provided the information-based tumour of above-mentioned rapid qualitative in one embodiment of the invention
The preparation method of former detection means, comprises the following steps：
1) labelled antibody is dissolved in buffer solution, the labelled antibody diluted, by the labelled antibody of described dilution
Glass fibre membrane is sprayed on, is then dried in vacuum desiccator, solid phase labelling antibody membrane is obtained；
2) coated antibody solution is lined and detection line is formed on reaction zone, rabbit anti-mouse igg solution is lined into reaction zone
Upper making nature controlling line formation check plot, the nitrocellulose film after line is placed under 37 DEG C of environment and dried 25 minutes, is coated with
Labelled antibody chromatographic film；
3) will be first in bottom plate from bottom to top paste sample uptake zone (the 1st area), filtering area (the 2nd area), chromatographic zone (the successively
3rd area), suction zones (the 7th area), paste makes the bottom that the overlapping 1~1.5mm in each area is assembled on viscose board, during stickup
After plate, the bottom plate of assembling is cut, the related specific antigen detector bar of tumour is obtained；
4) configuration instrument, network transmission device and terminal count device.
Below in conjunction with the embodiment in the present invention, the technical scheme in the present invention is clearly and completely described.It is aobvious
So, described embodiment is only a part of embodiment of the invention, rather than whole embodiments.Based on the reality in the present invention
Example is applied, the every other embodiment that those of ordinary skill in the art are obtained under the premise of creative work is not made all belongs to
In the scope of protection of the invention.
The configuration of the ovarian neoplasm specific antigen CA125 detection means of embodiment 1
1) setting of ovarian neoplasm specific antigen CA125 detector bars
By the ovarian neoplasm specific antigen CA125 labelled antibodies (JAJ that product type is #125-4012
International, Inc.San Diego, CA, USA) colloid gold label solution is added, after stirring 10~15 minutes, add dense
The Bovine serum albumin liquid for 1% is spent, the Bovine serum albumin liquid is 1 with colloid gold label liquor capacity ratio：7~15, warp
At a high speed after 4 DEG C of (12000PM) centrifugation 10 minutes, colored precipitate thing is obtained, the colored precipitate thing is labelled antibody.Blood in extraction
Clear liquid, sediment is dissolved in specific buffer solution.Labelled antibody mark after dilution is immersed on glass fibre membrane, will be soaked
Tunica fibrosa afterwards is under vacuum desiccator, and 35~38 DEG C are dried in vacuo, and solid phase labelling antibody membrane is made within 8 hours or so.Bag
It is 2mg/ml by the concentration of liquid；The coating buffer quantity for spray is to be sprayed 80ul/ seconds per 30cm nitrocelluloses film.By rabbit anti-mouse igg
The mg/ml speed of solution (product type #125-3401) 3 is the same, lines and nature controlling line C line (the 6th area) is made on reaction zone,
The check plot rule.Line is rule using lining instrument, model Biojet, the Biodot CA of the lining instrument,
USA.After line, nitrocellulose film is placed under 37 DEG C of environment and dried 25 minutes by the present invention, obtains labelled antibody chromatographic film.
Sample absorbing strip, filtering rod, mark chromatography strip and water suction bar are pasted onto on viscose glue bottom plate, the bottom assembled
Plate.Assemble method from bottom to top pastes sample uptake zone (the 1st area), filtering area (the 2nd area), chromatographic zone successively to be first in bottom plate
(the 3rd area), suction zones (the 7th area), paste makes the overlapping 1~1.5mm in each area on viscose board, during stickup.Assembled
Bottom plate after, the bottom plate of assembling is cut, slitting using cutting machine slitting, the cutting machine model K：
Nematic2360CA, USA.The width of the slitting is 4mm.Obtain ovarian neoplasm specific antigen CA125 detector bar.
2) in ovarian neoplasm specific antigen CA125 detection means instrument setting：
By mammary tumor CA-153 standard samples by following concentration dilution into negative serum, 0,35,70,140,280,
560 (u/ml units per mls).Above-mentioned 40ul microlitres (1 drop) is added in detector bar sample application zone, solvent 40ul is added immediately micro-
Rise (PBS), detection box is placed in instrument reading in 15 minutes, reading result summary sheet 3 is stored in detector
QikTech-4000 (JAJ international, Inc.San Diego, CA, USA) carries standby in software, and instrument is under
Table 2 obtains the color intensity (OD values) and mammary tumor CA-125 antigen concentration distribution curves in blood sample of precipitation line, and is stored in
In cloud server.
The different antigen concentration CA-125 of table 2 color intensity testing result
| CA125 concentration (u/ml units per mls) | Detected value 0.D |
| 0 | 0.02 |
| 35 | 0.10 |
| 70 | 0.21 |
| 140 | 0.54 |
| 280 | 0.92 |
| 560 | 1.62 |
3) in ovarian neoplasm specific antigen CA125 detection means network transmission device setting
Above-mentioned data are by the cloud service, and data receiver (such as inspection center doctor) is through wechat downloading data；
Or via detector QikTech-4000 output softwares, through USB output datas；Or it is defeated from wechat or the printing of bluetooth printing machine
Go out；Sent from the readout instrument of the detector by lettergram mode.
The configuration of the ovarian neoplasm specific antigen CA153 detection means of embodiment 2.
1) setting of ovarian neoplasm specific antigen CA153 detector bars：The preparation of ovarian neoplasm specific antigen CA153 detector bars
Be the same as Example 1 (prepare the antibody concentration that is used in detector bar and spray rate are), only by envelope antigen and labelled antigen
#153-782 and #153-678 are replaced with,
2) in ovarian neoplasm specific antigen CA153 detection means instrument setting：
Mammary tumor CA-153 standard samples are pressed into following concentration dilution into negative serum, 0,35,70,140,280,
560, (u/ml units per mls).Above-mentioned 40ul microlitres (1 drop) is added in detector bar sample application zone, solvent 40ul is added immediately
Microlitre (PBS), was placed in instrument reading, reading result summary sheet 3 is stored in detector in 15 minutes by detection box
QikTech-4000 (JAJ international, Inc.San Diego, CA, USA) carries standby in software, and instrument is under
Table obtains the color intensity (OD values) and mammary tumor CA-153 antigen concentration distribution curves in blood sample of precipitation line, and is stored in cloud
Hold in server.
The various concentrations CA-153 color intensity testing results of table 3
| CA153 concentration (u/ml units per mls) | Detected value 0.D |
| 0 | 0.08 |
| 35 | 0.17 |
| 70 | 0.31 |
| 140 | 0.58 |
| 280 | 0.94 |
| 560 | 1.70 |
3) in ovarian neoplasm specific antigen CA153 detection means network transmission device setting
Above-mentioned data are by the cloud service, and data receiver (such as inspection center doctor) is through wechat downloading data；
Or via detector QikTech-4000 output softwares, through USB output datas；Or it is defeated from wechat or the printing of bluetooth printing machine
Go out；Sent from the readout instrument of the detector by lettergram mode.
Embodiment 3
1) setting of ovarian neoplasm specific antigen CA199 detector bars：
The preparation be the same as Example 1 of ovarian neoplasm specific antigen CA153 detector bars (prepares the antibody concentration used in detector bar
Being with spray rate), envelope antigen and labelled antigen are only replaced with into #199-1033 and #199-1214,
2) in ovarian neoplasm specific antigen CA199 detection means instrument setting：
Mammary tumor CA199 standard samples are pressed into following concentration dilution into negative serum, 0,35,70,140,280,
560, (u/ml units per mls).Above-mentioned 40ul microlitres (1 drop) is added in detector bar sample application zone, solvent (phosphoric acid is added immediately
(PBS) buffer solution) 40ul microlitres, detection box is placed in instrument reading in 15 minutes, reading result summary sheet 3 is stored in
Detector QikTech-4000 (JAJ international, Inc.San Diego, CA, USA) carries standby in software, instrument
The color intensity (OD values) and mammary tumor CA-153 antigen concentration distribution curves in blood sample of precipitation line are obtained by following table, and is stored up
Exist in cloud server.
The various concentrations CA-199 testing results of table 4
| CA199 concentration (u/ml units per mls) | Detected value 0.D |
| 0 | 0.04 |
| 35 | 0.12 |
| 70 | 0.27 |
| 140 | 0.51 |
| 280 | 0.98 |
| 560 | 1.81 |
3) in ovarian neoplasm specific antigen CA199 detection means network transmission device setting
Above-mentioned data are by the cloud service, and data receiver (such as inspection center doctor) is through wechat downloading data；
Or via detector QikTech-4000 output softwares, through USB output datas；Or it is defeated from wechat or the printing of bluetooth printing machine
Go out；Sent from the readout instrument of the detector by lettergram mode.
The setting of the ED-SCLC specific antigen proGRP detection means of embodiment 4
1) setting of ED-SCLC specific antigen proGRP detector bars：
The preparation be the same as Example 1 of ED-SCLC specific antigen proGRP detector bars, it is only that envelope antigen is anti-with mark
Original replaces with #GRP-084 and #GRP-086,
2) in ED-SCLC specific antigen proGRP detection means instrument setting：
ED-SCLC proGRP standard samples are pressed into following concentration dilution into negative serum, 0,2.5,5.0,10,25,
50, (ng/ml units per mls).Add above-mentioned 40ul microlitres (1 drop) to give in detector bar sample application zone, solvent 40ul is added immediately
Microlitre (PBS), put quantitative instrument reading, reading result is converged, is stored in standby in detector software in 15 minutes by detection box.
The quantitative testing results of the ED-SCLC proGRP of table 5
| ProGRP(u/ml) | Color intensity |
| 0 | 0.061 |
| 2.5 | 0.242 |
| 5.0 | 0.401 |
| 10.0 | 0.812 |
| 25.0 | 1.594 |
| 50.0 | 2.602 |
3) in ED-SCLC specific antigen proGRP detection means network transmission device setting
Above-mentioned data are by the cloud service, and data receiver (such as inspection center doctor) is through wechat downloading data；
Or via detector QikTech-4000 output softwares, through USB output datas；Or it is defeated from wechat or the printing of bluetooth printing machine
Go out；Sent from the readout instrument of the detector by lettergram mode.
The setting of the ED-SCLC specific antigen Cyfra detection means of embodiment 4
1) setting of ED-SCLC specific antigen proGRP detector bars：
The preparation be the same as Example 1 of ED-SCLC specific antigen proGRP detector bars, it is only that envelope antigen is anti-with mark
Original replaces with #Cyf-124 and #Cyf-138,
2) in ED-SCLC specific antigen Cyffa detection means instrument setting：
Non-small cell lung cancer Cyffa21-1 standard samples are pressed into following concentration dilution into negative serum, 0,2.5,5.0,
10,25,50, (ng/ml units per mls).Add above-mentioned 40ul microlitres (1 drop) to give in detection box well, expansion is added immediately
40ul microlitres of agent (PBS), put quantitative instrument reading, reading result summary sheet 6 is stored in detector software in 15 minutes by detection box
In it is standby.
The quantitative testing results of the non-small cell lung cancer CYFRA21-1 of table 6
| Cyfra12-1(u/ml) | Color intensity |
| 0 | 0.079 |
| 2.5 | 0.257 |
| 5.0 | 0.419 |
| 10.0 | 0.745 |
| 25.0 | 1.615 |
| 50.0 | 2.670 |
3) in ED-SCLC specific antigen CYFRA21-1 detection means network transmission device setting
Above-mentioned data are by the cloud service, and data receiver (such as inspection center doctor) is through wechat downloading data；
Or via detector QikTech-4000 output softwares, through USB output datas；Or it is defeated from wechat or the printing of bluetooth printing machine
Go out；Sent from the readout instrument of the detector by lettergram mode.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of detection means of the related specific antigen of the information-based tumour of rapid qualitative, including the related specific antigen detection of tumour
Bar and instrument, network transmission device and terminal count device.
2. the detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claim 1, its feature exists
In the related specific antigen detector bar of the tumour includes sample uptake zone, filtering area, chromatographic zone and suction zones.
3. the detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claim 2, its feature exists
In the chromatographic zone includes immobilized label antibody district, detection zone and check plot.
4. the detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claim 1, its feature exists
In the instrument is handheld portable detector.
5. the detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claim 4, its feature exists
In the handheld portable detector is made up of the readout instrument based on smart mobile phone, inspection software, cloud service.
6. the detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claim 5, its feature exists
In the transmission means of the network transmission device includes the one or more of following methods：
By the cloud service, through wechat downloading data；
Via the detector output software, through USB output datas；
Printed out from wechat or bluetooth printing machine；
Sent from the readout instrument by lettergram mode.
7. the detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claim 5, its feature exists
In having stored the related specific antigen concentration of tumour and the detection of testing sample in the inspection software of the handheld portable detector
Linear relationship curve between the color intensity of instrument.
8. the detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claims 1 to 7, its feature
It is, the related specific antigen of the tumour includes mammary tumor specific antigen CA-125, mammary tumor specific antigen CA-153, breast
Room tumour specific antigen CA-199, ED-SCLC specific antigen proGRP, non-small cell lung cancer specific antigen Cyfra21-1
It is one or more of.
9. a kind of detection method of the related specific antigen of the information-based tumour of rapid qualitative, comprises the following steps：
1) sample that sample (blood, or serum, or blood plasma) is added dropwise into the specific antigen detector bar described in claim 1~7 adds
Sample area；
2) sample by or not by developping solution, into the filtering area described in claim 1~7, chromatographic zone；
If 3) there is " labelled antibody-antigen-solid-phase coating antibody " composite precipitation line in chromatographic zone, illustrate that testing result is
The positive, if there is not the composite precipitation line, illustrates to be detected as feminine gender；When the sample enters check plot, it should go out
Existing control line, when such as there is no control line, detection failure；
4) instrument described in application claim 1~7 is compound to " labelled antibody-antigen-solid-phase coating antibody "
Precipitation line carries out color intensity measure, obtains antigen concentration in sample, passes through the network transmission device described in claim 1~7
Transmit to the terminal count device.
10. a kind of detection means of the related specific antigen of the information-based tumour of rapid qualitative according to claims 1 to 7
Preparation method, it is characterised in that comprise the following steps：
1) labelled antibody is dissolved in buffer solution, the labelled antibody diluted, the labelled antibody of described dilution is sprayed
In glass fibre membrane, then it is dried in vacuum desiccator, obtains solid phase labelling antibody membrane；
2) coated antibody solution is lined and detection line is formed on reaction zone, rabbit anti-mouse igg solution is lined and made on reaction zone
Make nature controlling line formation check plot, the nitrocellulose film after line is placed under 37 DEG C of environment and dried 25 minutes, obtain coating mark
Antibody chromatographic film；
3) sample uptake zone, filtering area, chromatographic zone, suction zones first will be from bottom to top pasted successively in bottom plate, paste in viscose board
On after the bottom plate that is assembled, the bottom plate of assembling is cut, the related specific antigen detector bar of tumour is obtained；
4) configuration instrument, network transmission device and terminal count device.
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| CN110736830A (en) * | 2019-11-18 | 2020-01-31 | 威尚生物技术（合肥）有限公司 | detection cards for autoimmune colloidal gold |
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